LOCUS       sequence-210713-        9695 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GFP reporter of lag-2 driven by 405 bp of promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9695)
  AUTHORS   Pekar O, Ow MC, Hui KY, Noyes MB, Hall SE, Hubbard EJA
  TITLE     Linking the environment, DAF-7/TGFbeta signaling and LAG-2/DSL 
            ligand expression in the germline stem cell niche.
  JOURNAL   Development. 2017 Aug 15;144(16):2896-2906. doi: 10.1242/dev.147660.
  PUBMED    28811311
REFERENCE   2  (bases 1 to 9695)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: "10.1242/dev"; 
            journalName: "Development"; date: "2017-08-15- 15"; volume: "144"; 
            issue: "16"; pages: "2896-2906"
COMMENT     Sequence Label: pGC681
FEATURES             Location/Qualifiers
     source          1..9695
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1490..1501)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4559..4587)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     5335..5355
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5836..5853)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5836..5852)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7679..7701
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7739..7757)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7825..7929
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7930..7998,7999..8790)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGSDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8148..8167)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8961..9549
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9450..9469
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggccctgtcg gattatgggc tcttctttta tcctctactt tctttctaga aaatgcaaat
       61 gttgagtcag aaattgctga atggagtaca actgcaatgg gtttagctta tgtttctgga
      121 acaattactg gtggaattgt gattgaaaag taaggttata tttcatttca ataaatgtta
      181 ccacagtata acaaaaacaa gttacattgt tcagagtatg tctgaatgta ttattattat
      241 tgtatttgtg catcagtact gtagttgtac tgaagaaggg tttaaaataa tctcaacgtt
      301 tttaagacga gttagtgaga atacggtcgc agttaggatg tttggacata gttttattag
      361 ataatcgtga actaactcac tataaaaacc cgctaactca ttgaataaga tagttttaaa
      421 cgttcaaaat ttgaagacaa aattgtatga ataagtttac agagttggtc gacgcaagct
      481 tctccttctc ttcacatttc tcaataattt agctctaatt gcattcgtat tcttcgcaaa
      541 aatacgtatt ctgattgacc cgatgaaata cggatgccta gttgcattga tcatctacgg
      601 ttatacttat gggtgaggtt tttcaaatat attttgattt aatcgaataa tgaaatttca
      661 gaactggagt tggtccaatt tcgtggttta tatcctctga actggtacct caaaaacatc
      721 gtagtattgc tcaatctgtg gcatattcaa tgaatactgt aatggtggtt atcagtacat
      781 ttacagtatt acctctatat tccgttattg gtagttatgc tttccttatt ctatattcaa
      841 ttccttcatt tataagtatg cttatattat ttcgctactt accggaaacc aaaggacgag
      901 agattcatga tattgtgaat gaacttaaaa aaaaatgaaa acctttgaaa acgttcatac
      961 ggccgaaatt ttacttgatc gagccaattg aacttttgtt aagacttaca gaaaagtgag
     1021 agattttgtt acaaaaaagt gagaaaaaag cgaaaatgaa caggaaaaag aataaaaata
     1081 cacccgaagt atcatataaa aattaaaaag attgtttgac ctggcggaac ttttttgcaa
     1141 aaactctgaa tatccatggc acatgcttcg tgcaaaacag gatccaaatg aatatcaaca
     1201 agagattctt gaagacgacg agccacttgc tcagagcatt gcttatctcg aatgagaccc
     1261 ttgttaaaat catttttcaa acattatatg acattgtcga aatgtcctcc tgattccatg
     1321 atggtagcaa actcacttcg tctttgagcc aatttatcca agtccttgta aaaagtattc
     1381 gaaaattgtt aacggataaa atgtttatta taatatcaaa aacaatttgt cagttgacca
     1441 ctttttgata tatttgacag aaaccgggat gaattggctc aaagttaggg cgcccttcta
     1501 ttacagggtt tctgataaca aaccggttat taactcccaa caagggatga tttcaattca
     1561 tcatgctcaa attgacccaa attaagttac atgacaaatt catcgccctt ttcaactctt
     1621 cttggctcat catctgttat tctgttccta ttctgtctgc accctatacc cctttgcata
     1681 ctctctcgtc attcctttct tggatagtgt cttcttctcc ccagctctgc tacttctatg
     1741 acttgcgccc gctgcttttc cgctcgttcc tctcctctct gacgtcttct tctgcttctt
     1801 ttctccttcc tttttccctt ctcaatcttc ctctttttcc catttctgtc aatcattcga
     1861 agaagaagaa gaagaagacc ctcattcatt tcattttttt ttctgtcggt gtgtgctgct
     1921 cggttaagag tgagctctct attccacgtc ttcttctttt tcttcttcga ttcgaatcaa
     1981 tcactccacc aaaacgcatt cgtttttggg attcaccccg cggttcgcaa taggtttctt
     2041 tttcaaatta ttagcgttat aaatagaaaa atggtggagt ttcaataaaa atgataattt
     2101 acaaagtgta ttttgattac atgtactcaa aaaggttgaa atttttcata ccagttttcc
     2161 ggaaatccat ctgatatcat tatcgtattt ttctttttaa aaatgttttc aaaaaaaaac
     2221 aaaatatagc tggttatttg gcaccctcta attaccattt tcctgtcaca ccacactctt
     2281 ttccttcttc cacttctttc cgttttcagc cgcttccaac caaaccgata tgaaagccga
     2341 gcaacaacga tcgattccac ccggctcggc gaccttcccg tcgcaggtga gactcagaaa
     2401 actagagaaa ccgctcaact aactcttgat atccgatttc attcttttct cttttctttt
     2461 tgttgaactt ttccatattt ccagatgcca cggccaccac caagcaccga acaaggaatc
     2521 acaacggaat cggagcttgc gaagaaagct caaatcactc cgaacgacgt tttagcactt
     2581 ccgggaatca ctcaaggtat cttgctccct ttggatttca gtttaaaaca taaatttagg
     2641 attcttatgc tccccatctg cgaacatcta taacatcgag ttcaccaagt tccaaatccg
     2701 tgatctggac actgagcaag tgcttttcga gatcgccaaa ccggagaacg atcaggagaa
     2761 tgatgagtcg ccacaggagt cggcaagata cgtgcgttat agatttgctc caaacttttt
     2821 gaaactcaaa acggtcggag caactgtgga attcaaagta ggagacatcc caatccatca
     2881 tttccgaatg atcgaacgtc acttcttcaa agatcgcctt ctgaagtgtt ttgactttga
     2941 attcggattc tgtattccga attcacgaaa caactgtgaa catatctatg agttccctca
     3001 actctctcaa caactcagtg agtcattatt ctaaaaagta caattcaaaa gactaatctc
     3061 tttttcagtg gacgacatga tcaacaatcc aaacgagact cgttctgaca gcttctattt
     3121 cgtcgataac aaactcgtca tgcacaacaa agccgactac tcatatgatg cataaatatt
     3181 taatacaaaa atgttctgga taattattct gtcgaataga aaaaaaactc caaatgtgat
     3241 taaattccaa taattcctgt ctagtttgtt ccttcccctt ccccttctca tgttcaatgc
     3301 attcctaagc ttttcagttc cccccttgtt tctatatttt ttcgctgtcc tgtcacactc
     3361 gctaaaaaca ctaatcacac ggaaatctgt tttcaataaa aactccaact ttaactcatt
     3421 ttcaatttca actgaaagat tttttcatta gagaatgtct agaactaggc cgtttctctc
     3481 aagtattctt acacgtactt attttgtaca aatccttgtc agtcgctgca agaacataca
     3541 catcgaagag gcgcaatcga aacaacaggt gttcgcccgt tgatcgtctc cccgccccgc
     3601 ctgttgggcg ggacgggtgt cggtcaccac caccacatca tattgttgga cacactttgc
     3661 acatccggtt cacacccgat taccgcatcg gggtcttgat ctggggcggc tattggatct
     3721 tttgtttatg tagatttttt tctcgccgtt atgattttgg attatttttt ctcttatctt
     3781 gcagtcgact tgtaatcttt gcctgattgc tagctagcca aaactttcac ttgttctttt
     3841 ttttcctcta agtattttcc cgaacttata gtcgaccttg agctttgcta aagccaacaa
     3901 cacttttcaa atttgcccgc ggatccccat ctgcagaatt cgcccttaaa tgttcactcc
     3961 agaacagttc acaaaagtga tgtcccaact gggtaacttt tcccagttgg gacaaatgtt
     4021 ccaacccgat ccccgggatt ggccaaagga cccaaaggta tgtttcgaat gatactaaca
     4081 taacatagaa cattttcagg aggacccttg gagggtaccg gtccacgggc tccaggatga
     4141 cccggacctt caggcccttc tgaagggcag ccagcttctg aaggtgaagt ccagctcgtg
     4201 gcgtagggaa cgcttctaca agctacagga ggactgcaag accatctggc aggaatctcg
     4261 aaaggtcatg aggtccccgg agtcgcagct gttctccatc gaggacattc aggaggtacg
     4321 gatgggacac cgcacagaag gcctggagaa gtttgcccga gacatccccg aggatcgatg
     4381 cttctccatt gtcttcaagg accagcgcaa caccctagac ctcattgccc catcaccagc
     4441 tgacgctcag cactgggtgc agggcctgcg caagatcatc caccactccg gctccatgga
     4501 ccagcggcag aagggaccgg tagaaaaaat gagtaaagga gaagaacttt tcactggagt
     4561 tgtcccaatt cttgttgaat tagatggtga tgttaatggg cacaaatttt ctgtcagtgg
     4621 agagggtgaa ggtgatgcaa catacggaaa acttaccctt aaatttattt gcactactgg
     4681 aaaactacct gttccatggg taagtttaaa catatatata ctaactaacc ctgattattt
     4741 aaattttcag ccaacacttg tcactacttt ctgttatggt gttcaatgct tctcgagata
     4801 cccagatcat atgaaacggc atgacttttt caagagtgcc atgcccgaag gttatgtaca
     4861 ggaaagaact atatttttca aagatgacgg gaactacaag acacgtaagt ttaaacagtt
     4921 cggtactaac taaccataca tatttaaatt ttcaggtgct gaagtcaagt ttgaaggtga
     4981 tacccttgtt aatagaatcg agttaaaagg tattgatttt aaagaagatg gaaacattct
     5041 tggacacaaa ttggaataca actataactc acacaatgta tacatcatgg cagacaaaca
     5101 aaagaatgga atcaaagttg taagtttaaa catgatttta ctaactaact aatctgattt
     5161 aaattttcag aacttcaaaa ttagacacaa cattgaagat ggaagcgttc aactagcaga
     5221 ccattatcaa caaaatactc caattggcga tggccctgtc cttttaccag acaaccatta
     5281 cctgtccaca caatctgccc tttcgaaaga tcccaacgaa aagagagacc acatggtcct
     5341 tcttgagttt gtaacagctg ctgggattac acatggcatg gatgaactat acaaataagg
     5401 atgatcgacg ccaacgtcgt tgaattttca aattttaaat actgaatatt tgtttttttt
     5461 cctattattt atttattctc tttgtgtttt ttttcttgct ttctaaaaaa ttaattcaat
     5521 ccaaatctaa acattttttt ttctctttcc gtctcccaat tcgtattccg ctcctctcat
     5581 ctgaacacaa tgtgcaagtt tatttatctt ctcgctttca tttcattagg acgtgggggg
     5641 aattggtgga agggggaaac acacaaaagg atgatggaaa tgaaataagg acacacaata
     5701 tgcaacaaca ttcaattcag aaatatggag gaaggtttaa aagaaaacat aaaaatatat
     5761 agaggaggaa ggaaaactag taaaaaataa gcaaagaaat taggcgaacg atgagaattg
     5821 tcctcgcttg gaaacactgg ccgtcgtttt acactcgaga cgtacggtgc gcgcgatgca
     5881 ttcgaagatc tgcccactag tgagtcgtat tataagtgca agtaagatca gtgtttgttt
     5941 cgatatcagt ctgtttcgta acggtcttct gtataactac aaaaaagaat aaaaaaccgt
     6001 atcacaactg agaaaaaaga caaaacttat tgaacagtgt tcctttcaaa aaattatttt
     6061 gtaaaaaatc gagtttttct ccaaaataac gcacgcaatc aaaaacaaga aaactaacgt
     6121 gttctcctgg gcgtggaact ccttatcaag attaccatta ttgtctgtca gaattgatat
     6181 ttcaattatt tcatatctag acaaacccgg gtgtcttgca tttattagta ctgtgagttt
     6241 acatacaaac gaggcttact cagataacat agagtcacct ctaattagca aaaacccatt
     6301 tctaattagc tatttcgacc agttctaaag tcaccgtaat tagaaatggt ttttgtagct
     6361 tttctaatta gaaatgaccc cgagataagt gaaatatggc attttctggg ttttctagta
     6421 agaaatgatc ccgcacgtag tgattccgcg ttacaagcca aattcaaatt aaaaaagtat
     6481 attacacaat tcaagttaca ttattttcag gaaacaatgg cgaagagcaa agcagagatt
     6541 caaaaggagg aaagagacaa cagatatgaa cgaagacttc ttgaaaagag aaagagacat
     6601 gaggtgagta accatgataa taaataatca agtaagggct atagatttat aaattctgga
     6661 gttaacagtt tcgaaaaaaa gtagttttca tatgtgcgaa ggaatatcaa aaatagggga
     6721 atagcctaaa cgagtgagaa aattaaatgt ttgttatttt ttcaaaaatg tctgcagggc
     6781 cgttatcctt tcaaatgcac tctatcaaac atagatcata tccatcatat cttatccaga
     6841 atttatatca tatccaagat tgagagcgtt tcaaggaaaa tctcaaacaa ccatacaatt
     6901 ttgtagatcg acaacaaggc ctccacttct tcatcgatgg cctctgcagc acctgcacca
     6961 ccaagtttgt atttttcata ttaatataaa gtgagccacg catagtcgtt tttaggattt
     7021 ttgccagatc gatgtcgcgc aaacagactg tttttttttc tttgaaagcc gtgcgtgaca
     7081 tgggccacgg attgtaacgc cgcgctggat ggataaaaat ttattcagtt ttttagcctt
     7141 tacaaaatat ggttcgacta ctttctttat ttttacattt ttataggtgc tcattgagct
     7201 gagatatttt gggtataatc ggaaaatcgc tgattttggt catttttctg cactaaagat
     7261 tacacaaaaa gttcagtatt tagattcatt gttgaaaatt ggggatgatc tacagataca
     7321 tatcgccgtg ttcaaaaaca cacgtgctgt gacgtcgtga ctggggaaaa ccctggcgtt
     7381 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     7441 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg gcgcctgatg
     7501 cggtattttc tccttacgca tctgtgcggt atttcacacc gcatatggtg cactctcagt
     7561 gcaatctgct ctgatgccgc atagttaagc cagccccgac acccgccaac acccgctgac
     7621 gcgccctgac gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc
     7681 gggagctgca tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag acgaaagggc
     7741 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7801 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7861 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7921 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7981 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     8041 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     8101 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     8161 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     8221 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     8281 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     8341 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     8401 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     8461 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     8521 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8581 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8641 cgtgggtctc gcggtatcat tgcagcactg gggtcagatg gtaagccctc ccgtatcgta
     8701 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8761 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8821 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8881 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8941 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     9001 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     9061 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     9121 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     9181 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     9241 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     9301 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     9361 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     9421 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     9481 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     9541 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggtcttttg ctggcctttt
     9601 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     9661 gagtgagctg ataccagcgg ataacaattt cacag
//
